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Incomplete understanding of the root canal system leads to endodontic failure. Missed canal was the fourth most common endodontic failure, which needed retreatment. There were a few studies on internal morphology of posterior teeth of Myanmar population. However, there was no report on root canal anatomy of anterior teeth. So, the aim of the in vitro study was to investigate root canal anatomy of Myanmar permanent mandibular incisors in Mandalay Region by the staining and clearing method. A total of 96 teeth from the tertiary hospitals